Russian Journal of Genetics, Vol. 40, No. 3, 2004, pp. 271–281. Translated from Genetika, Vol. 40, No.

3, 2004, pp. 353–365.

Original Russian Text Copyright © 2004 by Chadov, Chadova, Kopyl, Khotskina, Fedorova.

GENERAL
GENETICS

Genes Controlling Development:

Morphoses, Phenocopies, Dimorphs,
and Other Visible Expressions of Mutant Genes
B. F. Chadov, E. V. Chadova, S. A. Kopyl, E. A. Khotskina, and N. B. Fedorova
Institute of Cytology and Genetics, Russian Academy of Sciences, Novosibirsk, 630090 Russia; e-mail: chadov@bionet.nsc.ru
Received May 22, 2003

Abstract—We studied facultative dominant lethal mutations obtained earlier in Drosophila melanogaster. In
some genotypes, these mutations were expressed as lethals, but in other genotypes they lacked this expression.
The mutations were maintained in the following cultures: (1) females Muller-5 heterozygous for the mutation;
(2) males crossed to attached-X females; and (3) females and males homozygous for the mutation. During cul-
turing, many mutations were found to give rise to phenotypically abnormal progeny. Generally, these abnor-
malities were morphoses involving various body parts; they were mostly asymmetric and nonheritable. Mater-
nal and paternal effects in the formation of morphoses were observed. In four cases, dimorphic mutations were
recorded: a female homozygous for the mutation had mutant phenotype whereas its male counterpart was phe-
notypically normal. The mutations were recessive with regard to the norm. New phenotypes behaving as muta-
tions with incomplete penetrance arose during culturing. In cultures of mutant homozygotes phenocopies
would appear en masse; they would persist for one or two generations and disappear. One wave of phenocopies
succeeded another. Visible phenotypes appeared, which further behaved as ordinary recessive mutations. We
concluded that these visible manifestations are characteristic for regulatory mutations controlling ontogeny.
Their appearance is explained by the activation of new regulatory scenarios caused by blocking standard regu-
latory pathways.

INTRODUCTION So far, we have developed three techniques of isola-

Any genic mutation is realized only in the process of
development. However, only some of these mutations
are involved in developmental control. Apparently,
these are mutations that concurrently possess two prop-
erties: they can arrest development and alter its progres-
sion. The former implies a particular importance of the
gene for the implementation of ontogeny, the latter con-
firms the regulatory nature of the gene.
These properties at first glance seem to be mutually
exclusive. However, this is not so. In 2000, an attempt
was made to find in Drosophila melanogaster a special
kind of mutation, which would be lethal in one geno-
type and nonlethal in another [1, 2]. The mutations of
this type were indeed found and termed facultative
dominant lethals. In some genotypes, such mutation
behaved as a dominant lethal, i.e., all the progeny bear-
ing this mutation died as early as at the embryonic
stage. In other genotypes, this mutation was not lethal,
i.e., the progeny were viable [3, 4].
Soon it was found that individuals with various
(sometimes severe) developmental malformations
occurred among the normal progeny of mutant parents
[4, 5]. Thus, the same mutation could either cause
lethality or disturb individual development. Mutations
having properties predicted for ontogeny-controlling
mutations in fact existed.
tion of facultative (also called conditional) lethals.
According to the first of them, a potential mutation is
obtained in the X chromosome of male D. melano-
gaster and then this male is crossed to yellow females.
The mutation is determined by the absence of female
progeny [1, 2]. The second technique is used to isolate
conditional dominant lethals in autosome 2 of D. mela-
nogaster [6]. The third method, like the first one, is
used for isolating mutations in the X chromosome, but
in the reverse order. First, lethal mutations are obtained
using the Muller-5 technique. These mutations mani-
fest their lethality in crosses between females
In(1)Muller-5/lethal with males In(1)Muller-5 (absence
of male lethal carriers in the progeny). Then, out of
these lethal mutations, the lethal are selected that do not
manifest lethality in crosses of the same females
In(1)Muller-5/lethal with males of genotypes different
from In(1)Muller-5 [7].
Lethality is the main property of these mutations:
they were detected by means of a test for lethality. In
the process of their maintenance in culture we have
found that their presence is manifested as the appear-
ance of morphoses in the progeny. However, morphoses
were not an only manifestation of these mutations. In
addition to morphoses, we have found (1) dimorphs,
i.e., mutant forms with mutation manifesting only in
one sex; (2) forms manifesting as visible mutations
with varying penetrance; (3) mass modifications; and

1022-7954/04/4003-0271 © 2004 MAIK “Nauka /Interperiodica”

272 CHADOV et al.
(4) forms manifesting as visible mutations with com-
plete penetrance. This article is devoted to phenotypic
description of the visible manifestation of the condi-
tional lethal mutations.
MATERIALS AND METHODS
Isolation of Mutations
Protocol 1. Males of D. melanogaster are exposed
to γ-irradiation and crossed to females carrying
attached-X chromosomes. Sons from this cross bear an
irradiated X chromosome and a haploid set of irradiated
autosomes. Each son is individually crossed to a yellow
female. The male that did not produce daughters is sup-
posed to carry an X-chromosomal mutation [1–3].
Protocol 2. Males of D. melanogaster are exposed
to γ-irradiation and crossed to females carrying inver-
sion Curly in autosome 2. Sons from this cross bearing
an irradiated autosome 2 and autosome Curly are indi-
vidually crossed to a yellow female. The male that did
not produce normal (not Curly) daughters and sons is
supposed to carry an autosome 2 mutation [3, 6].
Protocol 3. Recessive X-chromosomal lethals are
obtained in D. melanogaster using the classic Muller-5
method. The lethals are maintained by crossing
In(1)Muller-5/lethal females to In(1)Muller-5 males.
These crosses do not yield normal male progeny. In
crosses of the females with wild-type males, cultures
with appearing normal males are isolated. These cul-
tures contain the sought mutation [7].
Maintenance of Mutations
X-chromosomal mutations were maintained in
stock using two methods. In the first method, mutations
were carried by a female with a mutant X chromosome
(+) and an inverted chromosome Muller-5, B wa. In cul-
ture, these females were mated to males bearing a
mutant or inverted X chromosome. The progeny that
inherited the mutant chromosome from their mother
consisted of daughters +/B wa and wild-type sons (+).
The progeny that did not inherit the mutant chromo-
some from their mother consisted of daughters B wa/B wa
and sons B wa. In the second method, wild-type males
carrying the mutation were mated with females carry-
ing attached-X chromosomes (females C(1)DX, y w f/Y).
In these cultures, males inherit the mutant X chromo-
some paternally (father’s phenotype +) and daugh-
ters inherit attached-X chromosomes (mother’s phe-
notype y w f).
Mutations in autosome 2 were maintained in cul-
tures of flies that carried mutation in one autosome 2
and complex inversion In(2LR)CyO, in the other.
During culturing, we succeeded in isolating some of
the X-chromosomal mutations in a homozygous state.
In addition to the above methods, these mutations were
maintained in a homozygous state in males and
females.
RUSSIAN JOURNAL OF GENETICS
Examination of the cultures revealed novel phenotypes
[4, 5, 7], which were either stable or unstable. Using a
digital video camera, we obtained the pictures of the
mutant phenotypes and stored them as computer files.
RESULTS
Morphoses
Morphoses result from developmental disturbances
of different degree of severity, which do not prevent
eclosion, existence and mating of their carriers; these
flies even sometimes produce progeny.
Our collection contains several hundreds of colored
video pictures of morphoses, some of which are pre-
sented in Fig. 1. Morphological defects involved all
parts of the fly body and sometimes were extremely
severe (the absence of half of the head or thorax; one,
two, three or four legs; one wing; external genitalia).
Some of the new formations were similar to homeotic
ones: a doubled arista; the seventh leg; a tarsus on the
abdomen; protrusions on eyes, thorax, etc.; melanoma-
like neoplasms of various localization. The distribution of
morphoses over the fly body was presented earlier [4].
In contrast to visible mutations, morphoses (1) appear
in part of the progeny at a frequency varying from several
percent to several tens percent; (2) are not inherited;
(3) typically unilateral; (4) often put the individual at
the brink of extinction, but even in severe cases may
involve satisfactory viability and fertility.
In culturing X-chromosomal mutations, two types
of progeny appear: those bearing and not bearing the
mutant X chromosome. The morphose formation is
characterized by matro- and patrocliny. A morphose
may appear not only in the progeny that inherited the
mutant parental chromosome but also in the progeny
that did not. Obviously, the presence of the mutation in
a parent is a necessary condition for the morphose for-
mation in the progeny.
Maternal effect. Maternal effect appeared in the
progeny of females carrying a mutant X chromosome
(+) and an inverted chromosome Muller-5, B wa. In the
cultures, these females mated to males carrying a
mutant or inverted X chromosome. The progeny that
received a mutant chromosome from the mother, con-
sisted of daughters +/B wa and wild-type sons (+). The
progeny that had not received the mutant chromosome
consisted of daughters B wa/B wa and sons B wa. In a
group of seven X-chromosomal mutations, obtained in
a separate experiment (protocol 1), all mutants pro-
duced morphoses in their progeny. Five mutations
exhibited maternal effect on the morphose formation.
Twenty-six progeny of these mutants (daughters
B wa/B wa and sons B wa) showed malformations
despite the fact that they did not inherit a mutant
X chromosome from their mothers.
In a group of 24 mutations, isolated following pro-
tocol 3, morphoses were recorded in the progeny of all
mutant females Muller-5, B wa/+. Seventy-three (46%)
Vol. 40 No. 3 2004
 GENES CONTROLLING DEVELOPMENT 273

(a) (b)

(c) (d)

(e) (f)

(g) (h)

Fig. 1. Endogenous morphoses in D. melanogaster: (a) Sack-like protrusion on the median line of the lower thorax part; (b) absent
right wing, a necrosis focus at the wing base; (c) additional third metathoracic leg; (d) absent right thorax half; (e) cut right wing;
(f) absent femur and tibia part of the left metathoracic leg; (g) absent bristles and hair at the left thorax half, absent left wing;
(h) absent four legs, deformed leg and wing; (i) tarsus on the abdomen; (j) circular melanoma on the abdomen; (k) left head half
substituted by a modified arista; (l) head split in two; (m) rounded short right wing; (n) enlarged wing with a bubble at the right side;
(o) split thorax and disrupted abdominal tergite pattern; (p) additional wing as a protrusion on the right side.

out of 157 progeny with morphoses did not receive a
mutant X chromosome from their mothers. They
included 30 daughters and 43 sons. Thus, the formation
of a morphose in progeny and the presence in it of a
mutant chromosome are independent events. The for-
mation of morphoses without inheriting maternally a
mutant chromosome was recorded in special crosses of
mutant females +/Muller-5, B.
Paternal effect. A paternal effect was found in cul-
tures of attached-X females (females C(1)DX, y w f/Y)
and mutant wild-type males (+). In these cultures, sons

RUSSIAN JOURNAL OF GENETICS Vol. 40 No. 3 2004

274 CHADOV et al.

(i) (j)

(k) (l)

(m) (n)

(o) (p)

Fig. 1. (Contd.)

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 GENES CONTROLLING DEVELOPMENT
receive a mutant X chromosome from fathers, and
daughters receive attached X chromosome without
mutations from mothers. The paternal effect was
expressed in the appearance of daughter with morphoses.
In the group of seven mutations mentioned above, the
paternal effect manifested in four mutations (22 morpho-
ses). The paternal effect was observed also in crosses of
mutant males with other females carrying attached-X
chromosomes. These females were daughters with
morphoses, which did not receive any mutant X chro-
mosomes. In two mutations out of seven, both maternal
and paternal effects were observed.
In the group of 29 mutations isolated following pro-
tocol 1, which produced morphoses in the progeny, a
one-time examination of the progeny revealed morpho-
ses in the daughters. As noted above, the daughters did
not inherit mutant X chromosomes from their fathers.
The experimental data convincingly show that for the
mutations examined, the formation of pathological
phenotype (morphoses) without the inheritance of a
mutant gene is a rule rather than an exception.
Dimorphs: Mutation Expressed in One Sex
Several mutant-like forms having unusual, sex-
dependent expression appeared in the progeny of
mutants carrying conventional lethals. The mutational
nature of these forms is demonstrated by the possibility
to isolate them into cultures having 100 percent mani-
festation and stable inheritance in generations. We have
found four such mutations: short-legged, bubbly wing,
and two interrupted vein mutations (Fig. 2).
(1) short-legged (Fig. 2, top row). Males are pheno-
typically normal. In females, wings are obliquely cut
and dropped at sides. At eclosion of females, the upper
and lower blades of their wings are dissociated at the
median part and filled with hemolymph (“balloons”).
After hemolymph resorption, circular defects are left
on the wing. On both wings of females, longitudinal
vein L2 is interrupted (similar to a well-expressed
radius incompletus mutation). The tarsus is altered in
all six legs having one rather than five parts. This part
ends in two bristles. Females look short-legged because
of the absence of four tarsus parts. They are viable and
fertile but get easily stuck in the medium.
The mutation is not expressed in heterozygous
females. It was isolated shortly after finding using pro-
tocol 1. In cultures (females Muller-5/mutation × males
mutation and males Muller-5) homozygous B+ females
having this mutant phenotype appeared in addition to
heterozygous B/+ females. The mutation is maintained
in homozygous state by crossing with phenotypically
normal males. The penetrance in females is 100 percent.
Immediately after the isolation of the culture, 198 pheno-
typically normal sons and 168 short-legged daughters
in the scored progeny of 366 flies were observed. Dur-
ing a year of culturing, the appearance of one short-
legged male was recorded. This male left no progeny.
RUSSIAN JOURNAL OF GENETICS Vol. 40 No. 3
275
Mutant cultures with phenotypically normal homozy-
gous females were not served.
(2) bubbly wing (Fig. 2, bottom row). Males are phe-
notypically normal. Females have narrow wings with
upturned outer edge. At eclosion, balloons filled with
hemolymph occur on the median wing parts. After one
to two days, they disappear leaving circular defects
with impaired venation. A case of the appearance of a
male with the bubbly wing phenotype was recorded. We
failed to obtain a culture with manifestation of the char-
acter in both sexes.
The bubbly wing culture is maintained in a homozy-
gous and heterozygous variants. Heterozygous do not
exhibit the bubbly wing phenotype. A bubbly wing
female appeared in a culture (females Muller-5/muta-
tion × males mutation and males Muller-5), which con-
tained homozygous B/+ females with the above pheno-
type in addition to heterozygous B+ females.
In this culture, phenotypically normal females
occurred together with the bubbly wing females. The
former were used to obtain a sister culture, in which
both sexes are phenotypically normal.
Processes resulting in formation of a new phenotype
occur in the bubbly wing culture. Flies with impaired
tergites and short bristles appeared in it (Fig. 2b, bottom
row). This phenotype occurs in both sexes. Some flies
have small eyes (Fig. 2d, bottom row). The work on fix-
ing these new characters in a new derivative culture is
now in progress.
(3) Interrupted longitudinal vein (mutations 1 and 2).
Males are phenotypically normal. All females lack the
second longitudinal vein (radius incompletus, ri). The
size of the veinless area varies in progeny produced in
crosses with different strains but it is stable in the
mutant itself. Mutants carrying mutation 1 (Fig. 2b,
middle row) display only a short proximal fragment of
the vein, whereas in the case of mutation 2, the distal L2
fragment is preserved. The mutations are expressed
symmetrically on both wings. They are not expressed in
heterozygote. Females with impaired venation (muta-
tion 1) were detected immediately after obtaining
homozygotes for a conditional lethal. Out of 1205 prog-
eny, there was 450 ri daughters and 755 ri+ sons in the
dimorphic mutation 1 strain immediately after its isola-
tion. In subsequent scores carried out in four genera-
tions approximately six months after obtaining the
strain, yielded the counts of 126 ri daughters and
278 ri+ sons. Mutation ri no. 1 has a sister strain con-
taining the same lethal but lacking the interrupted lon-
gitudinal vein phenotype. All females and males in this
strain are phenotypically normal (ri+). During culturing
of the original strain Muller-5/mutation 1 (phenotype ri+),
two B/+ females with the interrupted longitudinal vein
phenotype. They founded a strain, in which both sexes
have visible interrupted longitudinal vein phenotype.
In one of derivative strains, penetrance of the ri pheno-
2004
276 CHADOV et al.

(a) (b) (c)

(a) (b) (c)

(a) (b) (c)

Fig. 2. Dimorphic mutations in D. melanogaster. Top row: mutation short-legged: (a) at right, a male without visible manifestation
of the mutation; at left, a female with the mutant phenotype (the absence of four tarsus members, obliquely cut, bubbly wings); (b) at
left, a heterozygous for the mutation female with normal 5-member tarsi; at right, a homozygous for the mutation female with short
tarsi; (c) total view of a female homozygous for the mutation. Middle row: mutation interrupted L2 vein: (a) a male without mani-
festation of the mutation; (b) absent L2 on both sides in a female homozygous for the mutation; (c) reduction of L4 and L5 in a male
(a new phenotype). Bottom row: mutation bubbly wings: at left, a male without the manifestation of the mutation; at right, a female
with “bubbly wings”; (b) new phenotype “defective tergites, short bristles” (male above, female below); (c) new phenotype “small
eyes,” a male).

type is complete; in another, it is lower, i.e., phenotyp-
ically normal flies appeared.
In the dimorphic strain, males with attenuated vein
L4 started to appear; then, this vein disappeared in pre-
viously phenotypically normal males (Fig. 2c, middle
row). Then we obtained females with the same pheno-
type. In one of the strains, the combination of pheno-
types is as follows: all females have the interrupted lon-
gitudinal vein phenotype, and all males, the interrupted
longitudinal vein together with the L4 phenotype. In
this case, we also observe sexual dimorphism but for
the character “absence of L4.”
In the case of dimorphic mutation ri (mutation 2),
homozygotes for the lethal had been for a long time
phenotypically normal. Then, two ri females appeared
among them; they were used to found the second
dimorphic ri culture. The expression of the mutation
was somewhat different: the gap in the vein was
smaller, and the distal vein fragment was present.
Unlike mutation 1, cultures no further phenotypic
change was observed in mutation 2.
The absence of the mutant phenotype in males was
not related to selective death of mutant males in the cul-
tures. The visible mutant manifestation in females was
recessive: females bearing the mutation in one X chro-
mosome and inversion Muller-5 in the other, were phe-
notypically normal.
Long-Term Modifications
Each of the obtained mutations has been maintained
in culture in five to ten vials. Each new generation is
scored under a binocular microscope to select parents
for the next generation.
The lethal mutations obtained were characterized by
phenotypic alterations of the modification type. This
was clearly seen in homozygous cultures, in which all

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 GENES CONTROLLING DEVELOPMENT
flies have the same normal phenotype. Scoring of each
new generation suddenly revealed up to ten flies with
altered phenotypes. We observed flies with dark-col-
ored body similar to those carrying the known black
mutation; light-colored body similar to yellow-2 pheno-
type (yellow cuticle, dark bristles), cut Notch-like
wings; altered form and order of abdominal tergites
(analogous to abnormal abdomen); reduced eyes (ana-
log of eyeless); obliquely cut dumpy-like wings. Waves
of the appearance of the brown phenotype (brown
instead of red eyes) were observed and massive forma-
tion of dark melanoma-like spots. These flies were indi-
vidually placed into vials to start new derivative cul-
tures. In most cases, the number of flies having the new
phenotype increased in the next generation. In deriva-
tive strains, the number of individuals with the new
phenotype was even higher. The expression level also
enhanced. However, after several generations, the num-
ber of flies having the new phenotype started to
decrease in both the original culture and the derivative
strain. Attempts to make the mutation homozygous
failed. In next “rounds,” flies with unusual phenotypes
generally ceased to appear. For instance, in the case of
“melanomas,” the penetrance of the character in the
derivative strain reached 100 percent. In spite of that,
flies with melanoma-like spots disappeared from the
cultures in three to four generations.
Flies with a special phenotype characteristically
appeared at once in several vials. The most enduring of
the modifications observed was biased sex ratio. In
some vials where the so-called “recessive sex-linked
lethal in the X chromosome” obtained by protocol 3
was maintained, males stopped to appear after one of
the transfers. The females to males ratio attained 200 : 1
and then, after four to five generations, returned to 1 : 1.
In some derivative strains, however, the sex ratio
remained biased: females outnumbered males approxi-
mately by a factor of 2.
Another remarkable event was the appearance of small
males in the cultures. These males were 2- to 2.5-fold
smaller in size than the normal males of the same
brood. The small males were fertile, but we failed to
obtain their culture though the number of their appear-
ance over all cultures was at least six.
One “wave” of the appearance of a particular modi-
fication could be succeeded by a wave of another mod-
ification, and then by a wave of still other one. Thus, the
wave of biased sex-ratio was followed by that of the
Notch-like phenotype (cuts at wing tips). The Notch-
like phenotype appeared at once in seven vials in differ-
ent derivative strains maintained for monitoring sex-
ratio change. The Notch-like flies form the progeny
were mated to one another. Only a few flies with this
phenotype were observed among several hundreds of
the progeny. In the next generation, no flies with the
Notch-like phenotype were found.
The receding Notch wave was followed by massive
appearance of males with the yellow-2 phenotype (yel-
RUSSIAN JOURNAL OF GENETICS Vol. 40 No. 3
277
low cuticle, dark bristles). About twenty males
appeared among several hundreds of the progeny. In the
next generation, no flies of either sex with the yellow-2
phenotype were found.
The appearance of mutant phenotypes described
here as modifications differed from that of morphoses.
Unlike morphoses, the appearance of a new phenotype
occurred massively and the phenotype was precisely
reproduced during several generations. Its expression
was the same on both the left and right sides of the
body. No severe and drastic, morphose-like distur-
bances were recorded. In general, the appearing pheno-
types imitated known mutations. They could be classi-
fied as phenocopies [8], but the latter are induced by
extraordinary external factors. In our case, they were
induced by the presence of a conditional lethal. Taking
into account the transmission of the mutant phenotypes
in generations, these phenotypic alterations should be
referred to as long-term modifications.
Visible Mutations with Incomplete Penetrance
Visible mutations with low penetrance often
appeared in the cultures of the lethal mutations. For
instance, a new ri mutation arose in the culture that pro-
duced the ri–ri+ dimorphic mutation. In contrast to the
dimorphic mutation, both males and females had the ri
phenotype but some of flies of either sex were pheno-
typically normal (ri+). The ri+ flies again produced ri+
and ri progeny. In six cultures homozygous for the con-
ditional lethal, we observed other abnormal phenotypes
caused by low-penetrance mutations. These visible
mutations were introduced in culture. Their phenotypes
are inherited according to the rules typical for low-pene-
trance mutations. Phenotypically they include tergite
defects (two abnormal abdomen-like variants), impaired
wing shape (two variants), brown eyes (of the brown
type), and “bubbly” wings.
Visible Mutations with Complete Penetrance
Visible mutations appeared in the cultures at a rate
higher than spontaneous but were not specifically stud-
ied. A derivative strain with such mutation could be eas-
ily obtained.
DISCUSSION
The problem of individual development, which is
clearly defined in biological studies, is not as clear
when addressed at the genetic level. In a sense, all or
nearly all genes are involved in ontogeny because all of
them function in the process of individual development.
Geneticists classify as ontogenetic genes controlling
development, such as homeotic genes [9, 10] or genes
of early development [11, 12]. There is yet no specific
criterion to distinguish among other genes those con-
trolling development.
2004
278 CHADOV et al.

(a) (b)

t4

t3

t2

t1

t0

Fig. 3. Genetic model of ontogeny. (a) Genes and signal pathways. The genome of an individual consists of structural genes (small
black circles) and regulatory genes of different ranks (large open circles, squares, and sectors). A regulatory ontogenetic gene is
represented by a set of cis-alleles (division of symbols into sectors). The genome is activated through a regulated system of signaling
pathways (lines between genes, arrows). The signaling pathways are terminated by switching of structural genes. Ontogeny is a pro-
cess of consecutive switching of regulatory genes of different rank according to the relay principle. A transition from the previous
to the next ontogenetic stage involves switching off of regulatory genes functioning at the former stage and structural genes provid-
ing the formation of presumptive structures (hatched circles); (b) ontogeny at a latter stage (t4). Switched off genes and inactive
signaling pathways are shown by a dotted line. Most structural genes and some regulatory genes with similar time of switching (reg-
ulatory genes of stem cells) are still in operation.

A discovery of regulatory genes [13] first in lower,
and then in higher organisms [14], motivated develop-
ment of ontogenetic schemes in the form of hierarchic
systems of regulatory genes. In these schemes, structural
genes are presented as completing elements [15–19].
However, the deployment of the developmental pro-
gram occurs in real live structures that require function-
ing of structural genes at all ontogenetic stages. To
depict the process of ontogeny in a systemic way, struc-
tural and regulatory genes should be combined in a log-
ical scheme.
In our graphic scheme of ontogeny (Fig. 3), struc-
tural genes are located at the ends of the regulatory
pathways. The main space is occupied by numerous of
hierarchically organized regulatory genes. Biological
characters are placed beyond the scheme outside the
structural genes. The characters are controlled by an
array of hierarchically connected regulatory genes,
which are ultimately meant to activate “in the right
place at the right time” particular structural genes sup-
plying specific cell families with their products [20].
In the proposed scheme, the development is shown
as a relay of activity passing through regulatory genes
of different rank. The signal pathways are terminated
by structural genes. Structural genes do not pass signals
but their activity is also of sequential wavelike charac-
ter. They could be regarded as developmental genes
both judging from their switching nature and from their
impact on development. However, in essence, they do
not control development. Mutations studied in the
present work affect regulatory genes.
The scheme shows why mutations of regulatory
developmental genes are lethals with incomplete pene-
trance. On the one hand, mutations of genes combined
in functional chains must arrest whole directions of
development, which causes lethality. On the other hand,
the lethal effect of the mutations can be suppressed
because of the existence of parallel signaling pathways.
Our scheme provides for this possibility. The regulatory
genes on it are divided in sectors. A possibility of exist-
ence of several rather than one link between the genes
is stipulated. These multiple links are not depicted in
the figure for the reasons of simplicity. The arguments
in favor of the complicated structure of regulatory
genes (cis-alleles) were discussed earlier [5].
The visible manifestation of lethals is an unexpected
property of genes responsible for individual develop-
ment. The forms of visible manifestation can be
arranged according to the degree of reproducibility of
the “visible phenotype”: in the immediate progeny and
in generations. The common visible mutations rank
first in this series; mutations with sex-dependent mani-
festation and visible mutations with varying expression
rank second and third, respectively; the fourth place in

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 GENES CONTROLLING DEVELOPMENT
the series is occupied by variations of non-mutation
nature: modifications and morphoses.
These forms of morphological variation have been
known to geneticists and described in a similar fashion.
The formation of morphoses commonly occurs when a
developing organism is exposed to extreme environ-
mental factors [21–25]. Visible mutations with low
penetrance have been known since classic studies of
Timofeev-Ressovsky on D. funebris [26–28]. Pheno-
copies and modifications have been described in many
organisms including Drosophila [8]. Mutations with
clearly dimorphic manifestations, apparently, are first
described in the present work although small sex-
dependent shifts in mutation expressions have been
repeatedly reported. However, the point is that all these
events have a common cause. They are all caused by
mutations of a particular category: conditional domi-
nant lethals. These mutations affect genes controlling
ontogeny.
Investigation of visible manifestation of ontogeny
genes is still restricted to observation. However, we can
fairly conclusively state that each of the mutations have
multiple manifestations. Each mutation is expressed at
least in three ways: as a (1) dominant lethal, (2) mor-
phose, and (3) no visible manifestation (norm). The
first and third form of manifestation allowed us to
detect the mutations. Morphoses in the progeny have
been recorded in the overwhelming majority of
mutants.
Such visible manifestations of mutations as mor-
phoses, modifications, and low-penetrance mutations
are characterized by low penetrance. Presumably, the
main property of ontogeny controlling genes is their
ability to assume two states: active and inactive. Since
these genes are of a paramount importance for develop-
ment, they must be present in multiple copies. This con-
clusion has been already reached earlier, as a result of
analysis of mutation lethality, and reflected in the regu-
latory gene model [5]. In the model, regulatory genes
are represented as cassettes of alleles in cis-position
(cis-alleles) [5]. Chromosome rearrangements and sex
can alter the activity of the cis-alleles [3, 5].
The visible gene manifestations confirm the conclu-
sion on the presence of the ontogeny genes in multiple
copies and on the activity alternation of the copies (cis-
alleles) in different individuals of the same species.
Bilateral asymmetry in morphose manifestation can be
also interpreted as resulting from multiple copies of the
ontogeny genes. One cis-allele (e.g., the normal one)
can act on the one side of the body while the other allele
(e.g., the mutant one) acts on the other side. As a result,
a morphose appears at the side of the mutant allele but
is absent at the other side of the body. This explanation
is in agreement with the general view on the asymmetry
mechanism advanced by Astaurov [29]. A strict deter-
mination of the regulatory gene order in a signal chain
seems to be slacker when selecting a cis-allele of a con-
crete gene.
RUSSIAN JOURNAL OF GENETICS Vol. 40 No. 3
279
From the theoretical viewpoint, the presence of mul-
tiple copies of ontogeny genes facilitates the construc-
tion of subprograms of development. Experimental
data indicate in Drosophila the existence of two such
subprograms: male and female programs of ontogeny.
The very fact of existence of dimorphic mutations tes-
tifies to this. The female ontogenetic subprogram
includes implementation of phenotypes “short-legged,”
“incomplete longitudinal vein,” and “bubbly wings.”
The male subprogram implements the normal variants
of these structures. We would like to emphasize that we
speak about phenotypic traits in Drosophila that do not
have sex distinctions. Because of this, the proposed
male and female programs of development are more
general than the existing concepts of sex determination
and differentiation [30] as a genetic basis of formation
of sex differences. We believe that these are separate
and independent programs of individual development.
The results on dimorphs indicate that the changes in
the female subprogram of the mutants were of a perma-
nent nature: each of the Drosophila females has a
mutant phenotype. However, no radical switch to
another female program occurred. The mutant pheno-
type was expressed in homozygous females but lacked
expression in heterozygous ones. Normally, the female
and male developmental programs are invariant: both
males and females keep their sexual appearance regard-
less of the presence of specific mutations. The change
of allele functioning in a dimorphic mutation occupies
an intermediate position between a normal (structural)
gene and a regulatory gene in the ontogenetic program.
Figure 4 presents a hypothetical scheme explaining
the existence of dimorphic mutations at regulatory
genes. Earlier it was suggested that in the normal pro-
gram of development, high-order regulatory genes func-
tion according to the principle of allele exclusion [5].
They are switched off in the opposite homolog (dashed
box in the figure). Normally, a signal is transmitted
from the cis-allele (d) at regulatory gene A to the cis-
allele (a) at gene B. In the case of dimorphic mutations,
in addition to the normal cis-allele (a), a mutant allele
(‡1) appears, which functions as the normal one. Con-
sequently, there are two variants of manifestation in
homozygote (a/a and ‡1/‡1) and one variant (‡1/‡), in
heterozygote. In the dimorphs, all three variants were
observed. A regulatory gene in principle works as a
structural gene, i.e., both trans-alleles function. For a
transition of the dimorphic manifestation of a regula-
tory gene to a monomorphic one (typical for a regula-
tory ontogenetic gene) requires “next to nothing”: sup-
pression of one of the two opposite trans-alleles in the pro-
cess of female ofr male gametogenesis (formation of a
non-alternative program of individual development [4]).
A transition of functioning of gene B from cis-allele
a to cis-allele b leads to the complete disappearance of
the mutant manifestation of the regulatory gene. This
phenomenon is specially considered for lethal expres-
sion of mutations [5]. If the transition is unstable, a typ-
2004
280 CHADOV et al.

A a1 B

a b c d a b c d

Fig. 4. Putative mechanism of operation of a dimorphic mutation. A and B are regulatory ontogenetic genes of the same rank. The
signal (horizontal arrow) passes from gene A to gene B. The downward chain of the regulatory genes of lower ranks leading to the
structural genes (black circles) is shown by a vertical arrow. Each of the regulatory genes is represented by a set of cis-alleles (small
squares). In the given organism, one cis-allele is functioning. Genes A and B function according to the rule of allele exclusion. Genes
of the top homolog are switched off (dotted boxes). Normally, the signal is transmitted along the (A)d–(B)a pathway. In the dimor-
phic mutant, both the normal (B) a and the mutant (B) ‡1 alleles are active. Thus, three allele combinations (‡/‡, ‡1/‡, and ‡1/‡1)
become possible.

ical mutation with incomplete penetrance appears. This
was characteristic for some derivatives of mutation ri
(incomplete longitudinal vein), which is a classic case
of incomplete penetrance of a visible mutation.
Judging from their relative frequency, the norm and
lethality are the most common forms of manifestation
of the mutations studied. Morphoses are rare. The fre-
quencies suggest that the normal development follows
a standard pathway upon signal transmission through
the chain of regulatory genes. A cis-allele mutation on
this pathway arrests development (is lethal). In rare
cases, the signal circumvents the hurdle, and the devel-
opmental arrest does not happen. However, in this case
the development is impaired due to a defect in the path-
way going down to the structural genes (Fig. 4, lateral
branches of the central pathway).
The collection of video images of various Droso-
phila morphoses that we obtained in screening the
mutants attains several hundred. The number of scored
morphoses is even higher. However, despite their diver-
sity, the number of morphose types seems to be
restricted. Some of them are more common and stan-
dard in appearance. One of the standard features is the
absence of a body part on one side of the fly; half tho-
rax, a wing, leg, haltere, or half tergite may be absent.
Apparently, as noted above, blocking the signal on a
standard pathway may result in its switching to another
pathway. The number of scenarios of abnormal devel-
opment is limited. A mutation in a regulatory develop-
mental gene provokes the expression of hidden devel-
opmental scenarios, which were formed in the organ-
ism but did not manifest. It suffices to compare the
existing variants of wing alterations (wing morphoses)
to be confident that they reflect definite developmental
variants of wing development rather than chaotic dis-
tortions of this organ.
The morphose formation occurred according to the
parental effect rule: the presence of a mutation in a par-
ent is sufficient for the appearance of a morphose in the
progeny. The presence of the mutation in the progeny
having the corresponding morphose was not obligatory.
Earlier, parental effect was reported for lethal expres-
sion of mutations [3–5]. This effect allowed us to for-
mulate a property, which principally distinguishes
genes of ontogeny from other regulatory genes. This
property is initiation of cell division by ontogeny genes.
An elementary ontogenetic event is “a transition from
one gene controlling ontogeny to another one through
cell division” [7]. Each step of the gene-to-gene transi-
tion of activity is backed up by the formation of cell
population of a strictly definite number. Thus is formed
and maintained the chain reaction of three intercon-
nected developmental processes: activation of genetic
information, augmentation of cell mass, and distribu-
tion of the activated information in the cell [7].

RUSSIAN JOURNAL OF GENETICS Vol. 40 No. 3 2004

 GENES CONTROLLING DEVELOPMENT
The main result of this study is a discovery of a
series of visible manifestations of genes controlling
ontogeny. The manifestation forms are diverse and
completely dissimilar to the form of mutation expres-
sion known from classic literature. It is likely that these
are manifestations of regulatory ontogenetic genes,
which is primarily indicated by the formation of mor-
phoses. The mechanisms of the formation of each man-
ifestation type are still vague. Genetic research of these
mutations is required.
ACKNOWLEDGMENTS
This work was supported by the Russian Foundation
for Basic Research, grant no. 01-01-48899.
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